Metabotropic glutamate receptors are associated with non-synaptic appendages of unipolar brush cells in rat cerebellar cortex and cochlear nuclear complex

Unipolar brush cells (UBCs) are a class of small neurons that are densely concentrated in the granular layers of the vestibulocerebellar cortex and dorsal cochlear nucleus. The UBCs form giant synapses with individual mossy fibre rosettes on the dendrioles which make up their brush formations and are provided with numerous, unusual non-synaptic appendages. In accord with the glutamatergic nature of mossy fibres, our previous post-embedding immunocytochemical studies indicated that various ionotropic glutamate receptor subunits are localized at the post-synaptic densities of the giant synapses, whereas the non-synaptic appendages are immunonegative. On the contrary, the metabotropic glutamate receptors mGluR1 and mGluR2/3 are situated at the non-synaptic appendages and are lacking at the post-synaptic densities. Other authors, however, have shown that antibodies to these metabotropic receptors stain both appendages and post-synaptic densities. In the present study, we have re-evaluated the distribution of metabotropic glutamate receptors in the UBCs of the cerebellum and the cochlear nuclear complex by light and electron microscopic pre-embedding immunocytochemistry with subtype-specific antibodies. We confirm that UBCs dendritic brushes are densely immunostained by antibody to mGluR1 particularly in the cerebellum and that antibody to mGluR2/3 labels at least a percentage of the UBC brushes in both the cerebellum and cochlear nuclei. At the ultrastructural level, it appears that mGluR1 and mGluR2/3 immunoreactivities are not associated with the post-synaptic densities of the giant mossy fibre–UBC synapses, but instead are concentrated on the non-synaptic appendages of the cerebellar UBCs. The non-synaptic appendages, therefore, may be an important avenue for regulating the excitability of UBCs and mediating glutamate effects on their still unknown intracellular signal transduction cascades. We also show that the pre-synaptic densities of UBC dendrodendritic junctions are mGluR2/3 positive. As previously demonstrated, antibodies to mGluR1 and mGluR2/3 label subsets of Golgi cells. Antibody to mGluR5 does not stain UBCs in the cerebellum and cochlear nucleus and reveals the somatodendritic compartment of Golgi cells situated in the core of the cerebellar granular layer, whilst cochlear nucleus Golgi cells are mGluR5 negative.     

9q34 loss of heterozygosity in a tuberous sclerosis astrocytoma suggests a growth suppressor-like activity also for the TSC1 gene

Tuberous sclerosis is an autosomal dominant disease whose characteristicfeature is the development of multiple hamartomas in a varietyof organs and tissues. Two major loci have been identified sofar: TSC1 on chromosome 9q34 and TSC2 on chromosome 16p13.3.Loss of heterozygosity at 16p13.3-associated markers has beenrecently observed in hamartomatous lesions of some tuberoussclerosis patients. Here we report the first evidence of lossof heterozygosity at the TSC1 critical region in a giant cellastrocytoma of a familial tuberous sclerosis case. Segregationanalysis showed that the 9q34 haplotype lost carried the putativenormal TSC1 gene. These data support the hypothesis of botha germline and somatic loss-of-function mutation for the developmentof tuberous sclerosis hamartomas and suggest a tumor-suppressor-likeactivity also for the TSC1 gene product. Finally, the possiblesignificance of a second small region of loss of heterozygosityat 9p21, found in the same astrocytoma, is discussed.     

Cellular uptake and metabolic reduction of pentavalent to trivalent arsenic as determinants of cytotoxicity and morphological transformation

Cytotoxicity, morphological neoplastic transformation, cellularuptake and metabolic reduction were determined in BALB/3T3 CIA31-1-1 cells for trivalent arsenic (sodium arsenite, As³⁺)and for pentavalent arsenic (sodium arsenate, As⁵⁺). The levelsof cellular uptake of⁷³As-labelled sodium arsenite and arsenatewere dose-dependent and highest in the first hour. At equimolarconcentration (3 x 10^–6 M), cellular uptake was 4-foldhigher for As³⁺ than for As⁵⁺. Cytotoxicity was higher for As³⁺than for As⁵⁺, but when correlated to total As cell burden itshowed no significant difference for the two forms. Morphologicaltransformation focus assays showed transforming activity forboth As³⁺ and As⁵⁺, with relative transformation frequenciesalso of 4:1. Recovery from the cytosol after exposure for 1–24h was >90% for either form of absorbed As. Exposure to As³⁺yielded 100% as As³⁺ in cytosol, but exposure to As⁵⁺ yielded>70% as As³⁺, showing a high rate of intracellular metabolicreduction. No methylated metabolites were detected by ion-exchangechromatography. After 24-h incubation in cell-free medium, oxidationof As³⁺ to As⁵⁺ occurred up to 30% of the dose, but incubationin the presence of cells lowered the oxidation level to 4%.As⁵⁺ was recovered unchanged from cell-free medium (24-h incubation),but in the presence of the cells it yielded up to 5% as As³⁺within 24 h and the cumulative release of As³⁺ by cells exposedto As⁵⁺ was dose-dependent. Glutathione depletion by diethylmaleateinhibited reduction of As⁵⁺ to As³⁺ by these cells up to 25%of controls, showing that As⁵⁺ reduction is partly dependenton glutathione. These results suggest that As³⁺ is the formresponsible for the cytotoxic and transforming effects, independentlyof the valence state of the inorganic arsenic in the culturemedium.     

Photo-reflex-rheographical method in the evaluation of elastocompression

The importance of a "compression-mobilization" association in order to attain a correct therapeutical procedure is stressed. The authors then report the results of a study conducted on patients affected by varices to establish the efficacy of a treatment by means of elastic stockings with graduated compressive pressure and the efficacy of reflex-rheography as a noninvasive, economical and easy-to-use method for the assessment of the variations in venous return flow produced by the association of muscular activity and elastocompression.     

Trans-Epithelial Transport,Metabolism, and Biological Activity Assessment of the Multi-Target Lupin Peptide LILPKHSDAD (P5) and Its Metabolite LPKHSDAD (P5-Met)

P5 (LILPKHSDAD) is a hypocholesterolemic peptide from lupin protein with a multi-target activity, since it inhibits both 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCoAR) and proprotein convertase subtilisin/kexin type-9 (PCSK9). This work shows that, during epithelial transport experiments, the metabolic transformation mediated by intestinal peptidases produces two main detected peptides, ILPKHSDAD (P5-frag) and LPKHSDAD (P5-met), and that both P5 and P5-met are linearly absorbed by differentiated human intestinal Caco-2 cells. Extensive comparative structural, biochemical, and cellular characterizations of P5-met and the parent peptide P5 demonstrate that both peptides have unique characteristics and share the same mechanisms of action. In fact, they exert an intrinsically multi-target behavior being able to regulate cholesterol metabolism by modulating different pathways. The results of this study also highlight the dynamic nature of bioactive peptides that may be modulated by the biological systems they get in contact with.     

Visual and Hearing Impairment Are Associated With Delirium in Hospitalized Patients: Results of a Multisite Prevalence Study

ObjectiveSensory deficits are important risk factors for delirium but have been investigated in single-center studies and single clinical settings. This multicenter study aims to evaluate the association between hearing and visual impairment or bi-sensory impairment (visual and hearing impairment) and delirium.DesignCross-sectional study nested in the 2017 “Delirium Day” project.Setting and ParticipantsPatients 65 years and older admitted to acute hospital medical wards, emergency departments, rehabilitation wards, nursing homes, and hospices in Italy.MethodsDelirium was assessed with the 4AT (a short tool for delirium assessment) and sensory deficits with a clinical evaluation. We assessed the association between delirium, hearing and visual impairment in multivariable logistic regression models, adjusting for: Model 1, we included predisposing factors for delirium (ie, dementia, weight loss and autonomy in the activities of daily living); Model 2, we added to Model 1 variables, which could be considered precipitating factors for delirium (ie, psychoactive drugs and urinary catheters).ResultsA total of 3038 patients were included; delirium prevalence was 25%. Patients with delirium had a higher prevalence of hearing impairment (30.5% vs 18%; P < .001), visual impairment (24.2% vs 15.7%; P < .01) and bi-sensory impairment (16.2% vs 7.5%) compared with those without delirium. In the multivariable logistic regression analysis, the presence of bi-sensory impairment was associated with delirium in Model 1 [odds ratio (OR) 1.5, confidence interval (CI) 1.2–2.1; P = .00] and in Model 2 (OR 1.4; CI 1.1–1.9; P = .02), whereas the presence of visual and hearing impairment alone was not associated with delirium either in Model 1 (OR 0.8; CI 0.6–1.2, P = .36; OR 1.1; CI 0.8–1.4; P = .42) or in Model 2 (OR 0.8, CI 0.6–1.2, P = .27; OR 1.1, CI 0.8–1.4, P = .63).Conclusions and implicationsOur findings support the importance of routine screening and specific interventions by a multidisciplinary team to implement optimal management of sensory impairments and hence prevention and the management of the patients with delirium.     

Normal sperm parameters per se do not reliably account for fertility: A case–control study in the real-life setting

A proportion of men are infertile despite having normal medical history/physical examination and normal semen analysis. We aimed to assess whether normal sperm parameters per se account for male factor fertility. 1,957 infertile men were compared with 103 age-comparable fertile controls. Semen analysis was based on 2010 World Health Organization reference criteria. Of all, 12.1% of infertile men and 40.8% of fertile men presented with normal sperm parameters. Among fertile men, 36.9% had isolated sperm abnormalities and 22.3% men showed two or more concomitant sperm abnormalities. Serum total testosterone was higher in infertile men with normal sperm parameters compared to those with ≥2 sperm abnormalities or azoospermia, but similar to those with isolated sperm abnormalities (p ≤ .001). Circulating hormones were similar among sperm parameters groups in fertile men. At multivariable analyses, testicular volume (OR 1.12, p ≤ .001) and FSH (OR 0.8, p ≤ .001) were associated with normal sperm parameters. Overall, the longer the infertility period, the greater the number of sperm parameters abnormalities (p < .01). In conclusion, we found that 12% of infertile men and only 41% of fertile men present with normal sperm parameters. Normal sperm parameters per se do not reliably account for fertility in the real-life setting.     

Conical Primary Cementless Stem in Revision Hip Arthroplasty: 94 Consecutive Implantations at a Mean Follow-Up of 12.7 years

BackgroundRevision of a failed total hip arthroplasty (THA) poses technical challenges. The use of primary stems for revision can be advantageous for maintaining bone stock and reducing complications: small case series have reported promising results in the short-term to mid-term follow-up. The aim of this study was to evaluate the long-term clinical and functional results and survivorship of a consecutive series of THA femoral component revisions using a conical primary cementless stem (PCS).MethodsNinety-four stem revisions with a preoperative Paprosky I or II defect were analyzed at an average follow-up of 12.7 ± 5.4 years. Aseptic loosening was the reason for revision in 92.5% of cases. Twenty patients were lost to follow-up. Two subgroups were created: Group 1 (n = 59) underwent isolated stem revision; Group 2 (n = 15) underwent complete THA revision. All were evaluated preoperatively and postoperatively based on the Harris Hip Score (HHS), the Western Ontario and McMaster Universities Index (WOMAC) score, and the visual analog scale for pain (VAS). Residual trochanteric pain and length discrepancies were recorded. Radiographic evaluation included signs of osteolysis, subsidence, loosening, and heterotopic ossification.ResultsPCS survivorship was 100% at 5 years and 95.9% at 10 years. Overall, significant postoperative improvements (P < .01) were observed on the HHS (44.3 vs 86.9), WOMAC (42.8 vs 82.8), and VAS (7.0 vs 3.0). Postoperative scores on all scales were higher for Group 1 (P < .01). Three patients (4.1%) underwent further stem revision. Demarcation lines (1 mm) were found in 12 (16.2%) patients and significant heterotopic ossifications in 22 (29.7%).ConclusionThe use of PCS for stem revision in failed THA with a limited femoral bone defect is a reliable option for both isolated stem revision and concomitant cup revision in well-selected patients.     

Liver transplantation versus liver resection for colorectal liver metastasis: a survival benefit analysis in patients stratified according to tumor burden score

Liver transplantation (LT) for colorectal liver metastasis (CRLM) may provide excellent survival rates in patients with unresectable disease. High tumor load is a risk factor for recurrence and low overall survival (OS) after liver resection (LR). We tested the hypothesis that LT could offer better survival than LR in patients with high tumor load. LR performed at Padua University Hospital for CRLM was compared with LT for unresectable CRLM performed both at Oslo and Padua. High tumor load was defined as tumor burden score (TBS) ≥ 9, and inclusion criteria were as in the SECA-I transplant study. 184 patients were eligible: 128 LRs and 56 LTs. 5-year OS after LR and LT was 40.5% and 54.7% (P = 0.102). In the high TBS cohort, 5-year OS after LR and LT was 22.7% and 52.2% (P = 0.055). In patients with Oslo score ≤ 2 and TBS ≥ 9 (13 LR; 24 LT) the 5-year OS after LR and LT was 14.6% and 69.1% (P = 0.002). The corresponding disease-free survival (DFS) was 0% and 22.9% (P = 0.005). Selected CRLM patients with low Oslo score and high TBS could benefit from LT with survival outcomes that are far better than what is achieved by LR.